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1.
Anesth Analg ; 132(1): 130-139, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-32167977

RESUMO

BACKGROUND: Intraoperative cardiac arrest (ICA) has a reported frequency of 1 in 10,000 anesthetics but has a much higher estimated incidence in orthotopic liver transplantation (OLT). Single-center studies of ICA in OLT are limited by small sample size that prohibits multivariable regression analysis of risks. METHODS: Utilizing data from 7 academic medical centers, we performed a retrospective, observational study of 5296 adult liver transplant recipients (18-80 years old) between 2000 and 2017 to identify the rate of ICA, associated risk factors, and outcomes. RESULTS: ICA occurred in 196 cases (3.7% 95% confidence interval [CI], 3.2-4.2) and mortality occurred in 62 patients (1.2%). The intraoperative mortality rate was 31.6% in patients who experienced ICA. In a multivariable generalized linear mixed model, ICA was associated with body mass index (BMI) <20 (odds ratio [OR]: 2.04, 95% CI, 1.05-3.98; P = .0386), BMI ≥40 (2.16 [1.12-4.19]; P = .022), Model for End-Stage Liver Disease (MELD) score: (MELD 30-39: 1.75 [1.09-2.79], P = .02; MELD ≥40: 2.73 [1.53-4.85], P = .001), postreperfusion syndrome (PRS) (3.83 [2.75-5.34], P < .001), living donors (2.13 [1.16-3.89], P = .014), and reoperation (1.87 [1.13-3.11], P = .015). Overall 30-day and 1-year mortality were 4.18% and 11.0%, respectively. After ICA, 30-day and 1-year mortality were 43.9% and 52%, respectively, compared to 2.6% and 9.3% without ICA. CONCLUSIONS: We established a 3.7% incidence of ICA and a 1.2% incidence of intraoperative mortality in liver transplantation and confirmed previously identified risk factors for ICA including BMI, MELD score, PRS, and reoperation and identified new risk factors including living donor and length of surgery in this multicenter retrospective cohort. ICA, while rare, is associated with high intraoperative mortality, and future research must focus on therapy to reduce the incidence of ICA.


Assuntos
Centros Médicos Acadêmicos/tendências , Parada Cardíaca/etiologia , Parada Cardíaca/mortalidade , Complicações Intraoperatórias/etiologia , Complicações Intraoperatórias/mortalidade , Transplante de Fígado/mortalidade , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Incidência , Transplante de Fígado/efeitos adversos , Masculino , Pessoa de Meia-Idade , Mortalidade/tendências , Estudos Retrospectivos , Fatores de Risco , Estados Unidos/epidemiologia , Adulto Jovem
2.
Microb Cell Fact ; 17(1): 17, 2018 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-29402276

RESUMO

BACKGROUND: Chimeric virus-like particles (VLP) allow the display of foreign antigens on their surface and have proved valuable in the development of safe subunit vaccines or drug delivery. However, finding an inexpensive production system and a VLP scaffold that allows stable incorporation of diverse, large foreign antigens are major challenges in this field. RESULTS: In this study, a versatile and cost-effective platform for chimeric VLP development was established. The membrane integral small surface protein (dS) of the duck hepatitis B virus was chosen as VLP scaffold and the industrially applied and safe yeast Hansenula polymorpha (syn. Pichia angusta, Ogataea polymorpha) as the heterologous expression host. Eight different, large molecular weight antigens of up to 412 amino acids derived from four animal-infecting viruses were genetically fused to the dS and recombinant production strains were isolated. In all cases, the fusion protein was well expressed and upon co-production with dS, chimeric VLP containing both proteins could be generated. Purification was accomplished by a downstream process adapted from the production of a recombinant hepatitis B VLP vaccine. Chimeric VLP were up to 95% pure on protein level and contained up to 33% fusion protein. Immunological data supported surface exposure of the foreign antigens on the native VLP. Approximately 40 mg of chimeric VLP per 100 g dry cell weight could be isolated. This is highly comparable to values reported for the optimized production of human hepatitis B VLP. Purified chimeric VLP were shown to be essentially stable for 6 months at 4 °C. CONCLUSIONS: The dS-based VLP scaffold tolerates the incorporation of a variety of large molecular weight foreign protein sequences. It is applicable for the display of highly immunogenic antigens originating from a variety of pathogens. The yeast-based production system allows cost-effective production that is not limited to small-scale fundamental research. Thus, the dS-based VLP platform is highly efficient for antigen presentation and should be considered in the development of future vaccines.


Assuntos
Apresentação de Antígeno , Pichia/genética , Vacinas de Partículas Semelhantes a Vírus/imunologia , Vacinas de Partículas Semelhantes a Vírus/isolamento & purificação , Animais , Patos , Hepatite B/imunologia , Antígenos de Superfície da Hepatite B/imunologia , Vírus da Hepatite B do Pato/imunologia , Humanos , Pichia/imunologia , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/isolamento & purificação , Vacinas Sintéticas/economia , Vacinas Sintéticas/imunologia , Vacinas de Partículas Semelhantes a Vírus/análise , Vacinas de Partículas Semelhantes a Vírus/genética
3.
J Sport Rehabil ; 27(2): 185-188, 2018 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-27992295

RESUMO

Clinical Scenario: Pain and range of motion (ROM) deficits are 2 issues that are commonly treated by clinicians. In certain instances, clinicians are tasked with treating patients who report with both pain and limited mobility. Currently, clinicians utilize a variety of different methods to combat pain and ROM limitations, but in singularity. However, contralateral exercises (CEs) may be a viable option that can have an effect on pain, ROM, or simultaneous effect on both. CLINICAL QUESTION: For patients with pain and/or ROM deficits, will CE decrease pain and increase ROM? SUMMARY OF FINDINGS: CE can have a significant effect on ipsilateral muscle activation, strength, as well as available motion on the contralateral limb. However, there is limited research on CE that explores effects on pain. Clinical Bottom Line: According to current evidence, CE can be a feasible option for clinicians trying to increase a patient's ROM. Furthermore, there can be enhanced effects on stability, muscle strength, and muscle activation due to CE. Strength of Recommendation: Studies that have been included are a level of 4 or higher based on Center for Evidence Based Medicine. However, future studies both of higher levels and variability should be conducted.


Assuntos
Terapia por Exercício/métodos , Manejo da Dor , Amplitude de Movimento Articular , Humanos , Força Muscular , Músculo Esquelético
4.
Anal Chem ; 86(16): 8368-75, 2014 Aug 19.
Artigo em Inglês | MEDLINE | ID: mdl-25066179

RESUMO

Microcantilever stress measurements are examined to contrast and compare their attributes with those from in situ X-ray absorption spectroscopy to elucidate bonding dynamics during the oxygen reduction reaction (ORR) on a Pt catalyst. The present work explores multiple atomistic catalyst properties that notably include features of the Pt-Pt bonding and changes in bond strains that occur upon exposure to O2 in the electrochemical environment. The alteration of the Pt electronic and physical structures due to O2 exposure occurs over a wide potential range (1.2 to 0.4 V vs normal hydrogen electrode), a range spanning potentials where Pt catalyzes the ORR to those where Pt-oxide forms and all ORR activity ceases. We show that Pt-Pt surface bond strains due to oxygen interactions with Pt-Pt bonds are discernible at macroscopic scales in cantilever-based bending measurements of Pt thin films under O2 and Ar. Complementary extended X-ray absorption fine structure (EXAFS) measurements of nanoscale Pt clusters supported on carbon provide an estimate of the magnitude and direction of the in-operando bond strains. The data show that under O2 the M-M bonds elongate as compared to an N2 atmosphere across a broad range of potentials and ORR rates, an interfacial bond expansion that falls within a range of 0.23 (±0.15)% to 0.40 (±0.20)%. The EXAFS-measured Pt-Pt bond strains correspond to a stress thickness and magnitude that is well matched to the predictions of a mechanics mode applied to experimentally determined data obtained via the cantilever bending method. The data provide new quantitative understandings of bonding dynamics that will need to be considered in theoretical treatments of ORR catalysis and substantiate the subpicometer resolution of electrochemically mediated bond strains detected on the macroscale.

5.
Microb Ecol ; 63(4): 761-72, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22202888

RESUMO

Pollutant effects on biofilm physiology are difficult to assess due to differential susceptibility of species and difficulty separating individual species for analysis. Also, measuring whole assemblage responses such as metabolism can mask species-specific responses, as some species may decrease and others increase metabolic activity. Physiological responses can add information to compositional data, and may be a more sensitive indicator of effect. It is difficult, however, to separate individual species for biochemical analyses. Agricultural runoff often contains multiple pollutants that may alter algal assemblages in receiving waters. It is unclear how mixtures containing potential algal growth stimulators and inhibitors (e.g., nutrients and herbicides) alter algal assemblage structure and function. In research presented here, algal biofilms were exposed to nutrients, atrazine, and their mixtures, and assemblage-level structural and functional changes were measured. Synchrotron infrared microspectroscopy (IMS) was used to isolate the biochemical changes within individual cells from a dominant species of a green alga (Mougeotia sp.), a diatom (Navicula sp.), and a cyanobacterium (Hapalosiphon sp.). At the assemblage level, mixtures generally increased algal biovolume, decreased chlorophyll a, and had no effect on metabolism or ammonium uptake. Navicula had a strong negative response to atrazine initially, but later was more affected by nutrients. Hapalosiphon responded positively to both atrazine and nutrients, and Mougeotia did not exhibit any biochemical trends. Generally, biochemical changes in each species were similar to cells experiencing low stress conditions, with increased relative protein and decreased relative lipid. IMS provided direct evidence that individual species in a natural biofilm can have unique responses to atrazine, nutrients, and mixtures. Results suggest that the initial benthic community composition should have a strong influence on the overall impact of agricultural pollutants.


Assuntos
Atrazina/farmacologia , Clorófitas/efeitos dos fármacos , Cianobactérias/metabolismo , Herbicidas/farmacologia , Poluentes Químicos da Água/farmacologia , Biofilmes/efeitos dos fármacos , Clorofila/metabolismo , Clorofila A , Clorófitas/metabolismo , Cianobactérias/efeitos dos fármacos , Diatomáceas/efeitos dos fármacos , Diatomáceas/metabolismo , Compostos de Amônio Quaternário/metabolismo , Espectrofotometria Infravermelho/métodos
6.
Front Immunol ; 12: 641421, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33815393

RESUMO

RTS,S is the leading malaria vaccine in development, but has demonstrated only moderate protective efficacy in clinical trials. RTS,S is a virus-like particle (VLP) that uses the human hepatitis B virus as scaffold to display the malaria sporozoite antigen, circumsporozoite protein (CSP). Particle formation requires four-fold excess scaffold antigen, and as a result, CSP represents only a small portion of the final vaccine construct. Alternative VLP or nanoparticle platforms that reduce the amount of scaffold antigen and increase the amount of the target CSP antigen present in particles may enhance vaccine immunogenicity and efficacy. Here, we describe the production and characterization of a novel VLP that uses the small surface antigen (dS) of duck hepatitis B virus to display CSP. The CSP-dS fusion protein successfully formed VLPs without the need for excess scaffold antigen, and thus CSP represented a larger portion of the vaccine construct. CSP-dS formed large particles approximately 31-74 nm in size and were confirmed to display CSP on the surface. CSP-dS VLPs were highly immunogenic in mice and induced antibodies to multiple regions of CSP, even when administered at a lower vaccine dosage. Vaccine-induced antibodies demonstrated relevant functional activities, including Fc-dependent interactions with complement and Fcγ-receptors, previously identified as important in malaria immunity. Further, vaccine-induced antibodies had similar properties (epitope-specificity and avidity) to monoclonal antibodies that are protective in mouse models. Our novel platform to produce VLPs without excess scaffold protein has wide implications for the future development of vaccines for malaria and other infectious diseases.


Assuntos
Imunogenicidade da Vacina/imunologia , Vacinas Antimaláricas/imunologia , Malária Falciparum/prevenção & controle , Proteínas de Protozoários/imunologia , Vacinas de Partículas Semelhantes a Vírus/imunologia , Animais , Anticorpos Antiprotozoários/imunologia , Malária Falciparum/imunologia , Camundongos , Plasmodium falciparum
7.
Transplant Direct ; 6(1): e520, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32047848

RESUMO

BACKGROUND: The assessment of perioperative risk factors for the development of acute respiratory distress syndrome (ARDS) has been described in various surgical populations. However, there are only limited data among patients undergoing liver transplantation (LT), particularly regarding the influence of intraoperative ventilation parameters. We sought to identify the perioperative risk factors associated with the development of ARDS in LT recipients. METHODS: This is a single-center, retrospective cohort study of adult patients who underwent LT at a tertiary academic medical center between January 1, 2006, and January 31, 2016. Postoperative ARDS was identified using the Berlin definition. Multivariable logistic regression analysis was used to identify perioperative risk factors for ARDS. RESULTS: Of 817 eligible patients who underwent an LT during the study period, 20 (2.45%) developed postoperative ARDS. In the preoperative model, ongoing dialysis (odds ratio, 6.41; P < 0.01) was identified as an independent risk factor of ARDS post-LT. A higher mean peak inspiratory pressure per increase of 1 cm H2O (odds ratio, 1.31; P < 0.01) was the only independent risk factor in the intraoperative model. Patients who developed ARDS postoperatively had significantly greater intensive care unit and hospital stay compared to non-ARDS patients (P < 0.001). There were no significant differences in the 30-day (P = 0.16) and 1-year (P = 0.51) mortality between the groups. CONCLUSIONS: Dialysis at the time of transplant and elevated intraoperative mean peak inspiratory pressure were associated with the development of ARDS. ARDS post LT was associated with increased intensive care unit and hospital length of stay, but not increased mortality.

8.
Analyst ; 134(6): 1099-106, 2009 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-19475135

RESUMO

Heart disease is by far the biggest killer in the United States, and type II diabetes, which affects 8% of the U.S. population, is on the rise. In many cases, the acute coronary syndrome and/or sudden cardiac death occurs without warning. Atherosclerosis has known behavioral, genetic and dietary risk factors. However, our laboratory studies with animal models and human post-mortem tissue using FT-IR microspectroscopy reveal the chemical microstructure within arteries and in the arterial walls themselves. These include spectra obtained from the aortas of ApoE-/- knockout mice on sucrose and normal diets showing lipid deposition in the former case. Also pre-aneurysm chemical images of knockout mouse aorta walls, and spectra of plaque excised from a living human patient are shown for comparison. In keeping with the theme of the SPEC 2008 conference 'Spectroscopic Diagnosis of Disease...' this paper describes the background and potential value of a new catheter-based system to provide in vivo biochemical analysis of plaque in human coronary arteries. We report the following: (1) results of FT-IR microspectroscopy on animal models of vascular disease to illustrate the localized chemical distinctions between pathological and normal tissue, (2) current diagnostic techniques used for risk assessment of patients with potential unstable coronary syndromes, and (3) the advantages and limitations of each of these techniques illustrated with patent care histories, related in the first person, by the physician coauthors. Note that the physician comments clarify the contribution of each diagnostic technique to imminent cardiac risk assessment in a clinical setting, leading to the appreciation of what localized intravascular chemical analysis can contribute as an add-on diagnostic tool. The quality of medical imaging has improved dramatically since the turn of the century. Among clinical non-invasive diagnostic tools, laboratory tests of body fluids, EKG, and physical examination are still the first line of defense. However, with the fidelity of 64-slice CT imaging, this technique has recently become an option when the patient presents with symptoms of reduced arterial flow. Single photon emission computerized tomography (SPECT) treadmill exercise testing is a standard non-invasive test for decreased perfusion of heart muscle, but is time consuming and not suited for emergent evaluation. Once the invasive clinical option of catherization is chosen, this provides the opportunity for intravascular ultrasound (IVUS) imaging. As the probe is pulled through the artery, the diameter at different parts is measurable, and monochrome contrast in the constricted area reveals the presence of tissue with a different ultrasonic response. Also, via an optical catheter with a fiber-optic conductor, the possibly of spectroscopic analysis of arterial walls is now a reality. In this case, the optical transducer is coupled to a near-infrared spectrometer. Revealing the arterial chemical health means that plaque vulnerability and imminent risk could be assessed by the physician. The classical emergency use of catherization involves a contrast agent and dynamic X-ray imaging to locate the constriction, determine its severity, and possibly perform angioplasty, and stent placement.


Assuntos
Vasos Sanguíneos/metabolismo , Cardiopatias/fisiopatologia , Fenômenos Ópticos , Idoso , Animais , Apolipoproteínas E/deficiência , Apolipoproteínas E/genética , Artérias/diagnóstico por imagem , Artérias/metabolismo , Vasos Sanguíneos/diagnóstico por imagem , Cateterismo , Angiografia Coronária , Doença da Artéria Coronariana/diagnóstico por imagem , Doença da Artéria Coronariana/metabolismo , Feminino , Técnicas de Inativação de Genes , Cardiopatias/diagnóstico por imagem , Cardiopatias/metabolismo , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Microespectrofotometria , Pessoa de Meia-Idade , Medição de Risco , Espectroscopia de Infravermelho com Transformada de Fourier , Fatores de Tempo , Tomografia Computadorizada por Raios X , Ultrassonografia de Intervenção
9.
J Biotechnol ; 306: 203-212, 2019 Dec 20.
Artigo em Inglês | MEDLINE | ID: mdl-31634510

RESUMO

Chimeric virus-like particles (VLP) are known as promising tools in the development of safe and effective subunit vaccines. Recently, a technology platform to produce VLP based on the small surface protein (dS) of the duck hepatitis B virus was established. In this study, chimeric VLP were investigated displaying the 195 N-terminal amino acids derived from the glycoprotein E2 of the bovine viral diarrhea virus (BVDV) on their surface. Isolation of the VLP from methylotrophic yeast Hansenula polymorpha was allowed upon co-expression of wild-type dS and a fusion protein composed of the BVDV-derived antigen N-terminally fused to the dS. It was shown the VLP could be purified by a process adapted from the production of a recombinant hepatitis B VLP vaccine. However, the process essentially depended on costly ultracentrifugation which is critical for low cost production. In novel process variants, this step was avoided after modification of the initial batch capture step, the introduction of a precipitation step and adjusting the ion exchange chromatography. The product yield could be improved by almost factor 8 to 93 ± 12 mg VLP protein per 100 g dry cell weight while keeping similar product purity and antigenicity. This allows scalable and cost efficient VLP production.


Assuntos
Vírus da Diarreia Viral Bovina/imunologia , Pichia/metabolismo , Vacinas de Partículas Semelhantes a Vírus/isolamento & purificação , Proteínas do Envelope Viral/metabolismo , Vacinas Virais/isolamento & purificação , Sequência de Aminoácidos , Proteínas do Capsídeo/genética , Vírus da Diarreia Viral Bovina/genética , Pichia/genética , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Vacinas de Partículas Semelhantes a Vírus/metabolismo , Proteínas do Envelope Viral/química , Proteínas do Envelope Viral/genética , Vacinas Virais/metabolismo
10.
PLoS One ; 14(9): e0221394, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31483818

RESUMO

BACKGROUND: Malaria caused by Plasmodium falciparum is one of the major threats to human health globally. Despite huge efforts in malaria control and eradication, highly effective vaccines are urgently needed, including vaccines that can block malaria transmission. Chimeric virus-like particles (VLP) have emerged as a promising strategy to develop new malaria vaccine candidates. METHODS: We developed yeast cell lines and processes for the expression of malaria transmission-blocking vaccine candidates Pfs25 and Pfs230 as VLP and VLP were analyzed for purity, size, protein incorporation rate and expression of malaria antigens. RESULTS: In this study, a novel platform for the display of Plasmodium falciparum antigens on chimeric VLP is presented. Leading transmission-blocking vaccine candidates Pfs25 and Pfs230 were genetically fused to the small surface protein (dS) of the duck hepatitis B virus (DHBV). The resulting fusion proteins were co-expressed in recombinant Hansenula polymorpha (syn. Pichia angusta, Ogataea polymorpha) strains along with the wild-type dS as the VLP scaffold protein. Through this strategy, chimeric VLP containing Pfs25 or the Pfs230-derived fragments Pfs230c or Pfs230D1M were purified. Up to 100 mg chimeric VLP were isolated from 100 g dry cell weight with a maximum protein purity of 90% on the protein level. Expression of the Pfs230D1M construct was more efficient than Pfs230c and enabled VLP with higher purity. VLP showed reactivity with transmission-blocking antibodies and supported the surface display of the malaria antigens on the native VLP. CONCLUSION: The incorporation of leading Plasmodium falciparum transmission-blocking antigens into the dS-based VLP scaffold is a promising novel strategy for their display on nano-scaled particles. Competitive processes for efficient production and purification were established in this study.


Assuntos
Antígenos de Protozoários/metabolismo , Vírus da Hepatite B do Pato/genética , Vacinas Antimaláricas/biossíntese , Pichia/metabolismo , Vacinas de Partículas Semelhantes a Vírus/biossíntese , Animais , Anticorpos Bloqueadores/imunologia , Antígenos de Protozoários/genética , Patos/virologia , Humanos , Malária/prevenção & controle , Vacinas Antimaláricas/imunologia , Vacinas Antimaláricas/isolamento & purificação , Plasmodium falciparum/metabolismo , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/isolamento & purificação , Vacinas de Partículas Semelhantes a Vírus/imunologia , Vacinas de Partículas Semelhantes a Vírus/isolamento & purificação
11.
PLoS One ; 14(9): e0221733, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31504038

RESUMO

The development of effective malaria vaccines remains a global health priority. Currently, the most advanced vaccine, known as RTS,S, has only shown modest efficacy in clinical trials. Thus, the development of more efficacious vaccines by improving the formulation of RTS,S for increased efficacy or to interrupt malaria transmission are urgently needed. The RTS,S vaccine is based on the presentation of a fragment of the sporozoite antigen on the surface of virus-like particles (VLPs) based on human hepatitis B virus (HBV). In this study, we have developed and evaluated a novel VLP platform based on duck HBV (known as Metavax) for malaria vaccine development. This platform can incorporate large and complex proteins into VLPs and is produced in a Hansenula cell line compatible with cGMP vaccine production. Here, we have established the expression of leading P. falciparum malaria vaccine candidates as VLPs. This includes Pfs230 and Pfs25, which are candidate transmission-blocking vaccine antigens. We demonstrated that the VLPs effectively induce antibodies to malaria vaccine candidates with minimal induction of antibodies to the duck-HBV scaffold antigen. Antibodies to Pfs230 also recognised native protein on the surface of gametocytes, and antibodies to both Pfs230 and Pfs25 demonstrated transmission-reducing activity in standard membrane feeding assays. These results establish the potential utility of this VLP platform for malaria vaccines, which may be suitable for the development of multi-component vaccines that achieve high vaccine efficacy and transmission-blocking immunity.


Assuntos
Vacinas Antimaláricas/imunologia , Proteínas de Protozoários/imunologia , Proteínas Recombinantes/imunologia , Vacinas de Partículas Semelhantes a Vírus/imunologia , Animais , Anopheles/parasitologia , Afinidade de Anticorpos , Células HEK293 , Vírus da Hepatite B/genética , Humanos , Vacinas Antimaláricas/genética , Mosquitos Vetores/parasitologia , Pichia/genética , Pichia/metabolismo , Plasmodium falciparum/genética , Plasmodium falciparum/imunologia , Plasmodium falciparum/patogenicidade , Proteínas de Protozoários/genética , Coelhos , Proteínas Recombinantes/genética , Vacinas de Partículas Semelhantes a Vírus/genética , Proteínas Virais/genética , Proteínas Virais/metabolismo
12.
Adv Biosyst ; 1(9)2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29552634

RESUMO

Complex 3D organizations of materials represent ubiquitous structural motifs found in the most sophisticated forms of matter, the most notable of which are in life-sustaining hierarchical structures found in biology, but where simpler examples also exist as dense multilayered constructs in high-performance electronics. Each class of system evinces specific enabling forms of assembly to establish their functional organization at length scales not dissimilar to tissue-level constructs. This study describes materials and means of assembly that extend and join these disparate systems-schemes for the functional integration of soft and biological materials with synthetic 3D microscale, open frameworks that can leverage the most advanced forms of multilayer electronic technologies, including device-grade semiconductors such as monocrystalline silicon. Cellular migration behaviors, temporal dependencies of their growth, and contact guidance cues provided by the nonplanarity of these frameworks illustrate design criteria useful for their functional integration with living matter (e.g., NIH 3T3 fibroblast and primary rat dorsal root ganglion cell cultures).

14.
J Biotechnol ; 225: 3-9, 2016 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-26988393

RESUMO

Streptavidin - a protein secreted by the filamentous bacterium Streptomyces avidinii - is applied in a variety of methods, leading to numerous studies on its heterologous production. Development and characterization of a novel expression system for streptavidin genes by Hansenula polymorpha is described utilizing different target gene variants along with the two methanol-inducible promoters PMOX and PFMD. Extracellular product concentrations were higher for cultivation at 30 instead of 37°C. The best performing strain carrying the full-length streptavidin gene under control of PFMD was characterized in the bioreactor applying a synthetic medium and oxygen-controlled feeding of glucose. Derepression resulted in an extracellular concentration of 1.31±0.07µM of tetrameric streptavidin after 48h (27.3nMh(-1)). Feeding of glycerol improved biomass formation, but lowered the product concentration. By combining derepression and methanol induction the final extracellular streptavidin concentration increased to 11.42±0.22µM (approx. 751mgL(-1)), yielding a productivity of 52.5nMh(-1). Despite supplementing biotin the proportion of biotin-blocked binding sites in the supernatant dropped from 54.4±5.0 % after 18h to 17.2±6.5 % towards the end of glucose feeding to a final value of 1.1±3.8 %, indicating a highly bioactive product. Thus, H. polymorpha proved to be a suitable host for the production of streptavidin.


Assuntos
Pichia/crescimento & desenvolvimento , Regiões Promotoras Genéticas , Estreptavidina/genética , Estreptavidina/metabolismo , Técnicas de Cultura Celular por Lotes , Biomassa , Meios de Cultura/química , Meios de Cultura/farmacologia , Fermentação , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Glicerol/química , Metanol/farmacologia , Pichia/genética
15.
J Biotechnol ; 221: 91-100, 2016 Mar 10.
Artigo em Inglês | MEDLINE | ID: mdl-26820322

RESUMO

Due to its various applications the protein streptavidin is a highly interesting target for heterologous production. This study focuses on different Escherichia coli-based constructs targeting a high-level expression and secretion of streptavidin to the medium. The effect of various promoters, variants of the target gene, leader sequences and host strains on expression and secretion into the culture broth was analyzed. Constitutive production of full-length streptavidin fused with the leader sequence of the bglA gene from Bacillus amyloliquefaciens by the periplasmic 'leaky mutant' E. coli JW1667-5 (Δlpp-752:kan) at 30°C generated the highest yield of the conditions tested, surpassing the extracellular concentration of a conventional T7-based expression system. Supplementation of the medium by the non-ionic surfactants Triton(®) X-100 and X-45 led to an improved secretion of the protein to the culture supernatant. Tetrameric concentrations of streptavidin of 2790±166nM were reached in shake flasks at a productivity of 49.6nMh(-1). Optimization of conditions led to a successful transfer to the bioreactor, yielding a maximal concentration of 2608±169nM and a productivity of 65.2nMh(-1) in fed-batch operation. The proportion of biotin-blocked binding sites of 8.3±4.3% indicated a highly bioactive product.


Assuntos
Bacillus/genética , Escherichia coli/crescimento & desenvolvimento , Mutação , Estreptavidina/biossíntese , Regiões 5' não Traduzidas , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Técnicas de Cultura Celular por Lotes , Clonagem Molecular , Meios de Cultura/química , Escherichia coli/genética , Regiões Promotoras Genéticas , Estreptavidina/genética , Estreptavidina/metabolismo
16.
Adv Mater ; 28(4): 742-7, 2016 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-26618617

RESUMO

A 3D mechanically stable scaffold is shown to accommodate the volume change of a high-specific-capacity nickel-tin nanocomposite during operation as a Li-ion battery anode. The nickel-tin anode is supported by an electrochemically inactive conductive scaffold with an engineered free volume and controlled characteristic dimensions, which engender the electrode with significantly improved cyclability.

17.
Adv Healthc Mater ; 5(9): 1025-39, 2016 05.
Artigo em Inglês | MEDLINE | ID: mdl-26924676

RESUMO

3D hydrogel scaffolds are widely used in cellular microcultures and tissue engineering. Using direct ink writing, microperiodic poly(2-hydroxyethyl-methacrylate) (pHEMA) scaffolds are created that are then printed, cured, and modified by absorbing 30 kDa protein poly-l-lysine (PLL) to render them biocompliant in model NIH/3T3 fibroblast and MC3T3-E1 preosteoblast cell cultures. Spatial light interference microscopy (SLIM) live cell imaging studies are carried out to quantify cellular motilities for each cell type, substrate, and surface treatment of interest. 3D scaffold mechanics is investigated using atomic force microscopy (AFM), while their absorption kinetics are determined by confocal fluorescence microscopy (CFM) for a series of hydrated hydrogel films prepared from prepolymers with different homopolymer-to-monomer (Mr ) ratios. The observations reveal that the inks with higher Mr values yield relatively more open-mesh gels due to a lower degree of entanglement. The biocompatibility of printed hydrogel scaffolds can be controlled by both PLL content and hydrogel mesh properties.


Assuntos
Fibroblastos/metabolismo , Hidrogéis/química , Poli-Hidroxietil Metacrilato/química , Polilisina/química , Alicerces Teciduais/química , Animais , Camundongos , Células NIH 3T3
18.
ACS Appl Mater Interfaces ; 7(33): 18406-14, 2015 Aug 26.
Artigo em Inglês | MEDLINE | ID: mdl-26258957

RESUMO

Although rechargeable magnesium (Mg) batteries show promise for use as a next generation technology for high-density energy storage, little is known about the Mg anode solid electrolyte interphase and its implications for the performance and durability of a Mg-based battery. We explore in this report passivation effects engendered during the electrochemical cycling of a bulk Mg anode, characterizing their influences during metal deposition and dissolution in a simple, nonaqueous, Grignard electrolyte solution (ethylmagnesium bromide, EtMgBr, in tetrahydrofuran). Scanning electron microscopy images of Mg foil working electrodes after electrochemical polarization to dissolution potentials show the formation of corrosion pits. The pit densities so evidenced are markedly potential-dependent. When the Mg working electrode is cycled both potentiostatically and galvanostatically in EtMgBr these pits, formed due to passive layer breakdown, act as the foci for subsequent electrochemical activity. Detailed microscopy, diffraction, and spectroscopic data show that further passivation and corrosion results in the anisotropic stripping of the Mg {0001} plane, leaving thin oxide-comprising passivated side wall structures that demark the {0001} fiber texture of the etched Mg grains. Upon long-term cycling, oxide side walls formed due to the pronounced crystallographic anisotropy of the anodic stripping processes, leading to complex overlay anisotropic, columnar structures, exceeding 50 µm in height. The passive responses mediating the growth of these structures appear to be an intrinsic feature of the electrochemical growth and dissolution of Mg using this electrolyte.

19.
Mayo Clin Proc ; 88(7): 674-82, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23809318

RESUMO

OBJECTIVE: To describe the perioperative management of patients with left ventricular assist devices (LVADs) who require general anesthesia while undergoing noncardiac surgery (NCS) at a single, large tertiary referral center. PATIENTS AND METHODS: Electronic medical records from September 2, 2005, through May 31, 2012, were retrospectively reviewed to evaluate the perioperative management and outcomes in LVAD patients undergoing NCS. Patients were included only if they required a general anesthetic and had previously been discharged from the hospital after initial LVAD implantation. RESULTS: Thirty-three patients with LVADs underwent general anesthesia for 67 noncardiac operations. The mean ± SD time from LVAD implantation to NCS was 317 ± 349 days. All but 1 patient had axial flow LVADs. Anticoagulation or antiplatelet agents were present within 7 days before NCS in 49 procedures (73%) and reversed in 32 of 49 (65%). No perioperative thrombotic complications related to anticoagulation or antiplatelet reversal were noted. Red blood cell, fresh frozen plasma, and platelet transfusions were administered during 10, 6, and 4 operations, respectively. The only intraoperative complication was surgical bleeding. Postoperative complications were present in 12 patients after NCS and were mainly composed of bleeding. Three patients died within 30 days of NCS, with the causes of death not attributed to NCS. CONCLUSION: Patients with LVAD safely underwent NCS in a multidisciplinary setting that included preoperative optimization by cardiologists familiar with LVADs when feasible. Anticoagulation or antiplatelet agents were present preoperatively in most patients with LVADs and were safely reversed when necessary for NCS. The relatively high occurrence of postoperative bleeding is consistent with previous series.


Assuntos
Coração Auxiliar/estatística & dados numéricos , Assistência Perioperatória/estatística & dados numéricos , Complicações Pós-Operatórias/epidemiologia , Procedimentos Cirúrgicos Operatórios/estatística & dados numéricos , Disfunção Ventricular Esquerda/epidemiologia , Disfunção Ventricular Esquerda/terapia , Idoso , Anestesia Geral/estatística & dados numéricos , Causalidade , Comorbidade , Feminino , Hemorragia/etiologia , Humanos , Masculino , Pessoa de Meia-Idade , Cuidados Pós-Operatórios/estatística & dados numéricos , Cuidados Pré-Operatórios/estatística & dados numéricos , Estudos Retrospectivos , Índice de Gravidade de Doença , Procedimentos Cirúrgicos Operatórios/efeitos adversos , Resultado do Tratamento
20.
Appl Spectrosc ; 64(12): 1320-4, 2010 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21144147

RESUMO

In the dry milling of wheat flour, each unit process (roller mill, purifier, sifter, etc.) produces a mixture with varying amounts of wheat endosperm and non-endosperm byproducts. Chemical images with 82 000 pixels of each intermediate product stream issuing from an individual processing machine are readily analyzed in terms of the relative amount of endosperm and non-endosperm. Approximately three minutes is required to produce an image of each intermediate product stream. Applying partial least squares (PLS) chemometric software to identify individual pixels, which enables calculation of the relative amount of endosperm and non-endosperm, is not a time-limiting factor. When relative flow rates are known for each stream, mass balance can be calculated from each intermediate stream in terms of the product (endosperm content) and the lower value non-endosperm byproduct. Data is presented from a purifier in a commercial flour mill. Intermediate streams collected from a run with optimized operational parameters were compared to those of another run before adjustment. The endosperm (product) mass balance profile for each run enabled assessment of operational efficiency. The devised chemical imaging analysis system would be particularly useful in commissioning of a new mill or to optimize existing wheat milling systems. Also, when raw material differs from that for which previous optimization was established, a new optimization may be in order. The ability to acquire a large number of spectra from a specimen and apply multivariate statistics to identify each pixel and subsequently count pixels accommodates heterogeneity and reports the results from averaging a very large number of individual spectra. A second illustration of the utility of the imaging method is presented centering on streams from the first and second break unit operations at the beginning of the roller mill process.


Assuntos
Endosperma/química , Farinha/análise , Manipulação de Alimentos/métodos , Espectroscopia de Luz Próxima ao Infravermelho/métodos , Triticum/química , Análise dos Mínimos Quadrados
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